RESUMO
The results on in vitro culture establishment, plantlet regeneration and rooting of Camelina sativa cultivar sample Peremozhets and cultivar Mirazh are presented. Effective concentrations of sterilizing agents and duration of plant material treatment were estimated. Phytohormone ratio, sucrose concentration in nutrient medium that induce effective formation of C. sativa shoots and NAA concentration for plantlet rooting have been established. The method of Agrobacterium-mediated transformation of Camelina by using binary vector pGH217 carrying reporter beta-glucoronidase (gus) gene driven under 35S CaMV promoter and nos-terminator, and selective marker hpt gene conferring hygromycin-resistance in transgenic plant was elaborated.
Assuntos
Brassicaceae/crescimento & desenvolvimento , Brotos de Planta/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Regeneração/efeitos dos fármacos , Sementes/crescimento & desenvolvimento , Transformação Genética/efeitos dos fármacos , Agrobacterium/genética , Compostos de Benzil/farmacologia , Brassicaceae/genética , Brassicaceae/fisiologia , Cinamatos/farmacologia , Técnicas de Cocultura , Glucuronidase/genética , Higromicina B/análogos & derivados , Higromicina B/farmacologia , Ácidos Naftalenoacéticos/farmacologia , Reguladores de Crescimento de Plantas/farmacologia , Brotos de Planta/genética , Brotos de Planta/fisiologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Purinas/farmacologia , Sementes/genética , Sementes/fisiologiaRESUMO
The data of Agrobacterium-mediated transformation of some Linum usitatissimum cultivars zoned on the territories of Belarus and Ukraine with the plasmid carrying chimeric GFP-TUA6 gene and nptII gene as selectable marker conferring resistance to kanamycin are presented in this study. Transformation was affected by a number of factors including optical density (OD600), time of inoculation of explants with Agrobacterium and co-culture conditions. Transgenic nature of obtained lines was confirmed by PCR analysis. Expression of GFP-TUA6 gene was detected with confocal laser scanning microscopy. The obtained transgenic lines can be used for further functional studies the role of microtubules in the processes of building the flax fibres and resistance to wind.
Assuntos
Linho , Proteínas de Fluorescência Verde/genética , Microtúbulos/ultraestrutura , Proteínas Recombinantes de Fusão/genética , Transformação Genética , Tubulina (Proteína)/genética , Agrobacterium tumefaciens/genética , Proteínas de Arabidopsis/genética , DNA de Plantas/genética , Linho/genética , Linho/ultraestrutura , Genes de Plantas , Microscopia Confocal , Microtúbulos/genética , Plantas Geneticamente Modificadas , Reação em Cadeia da PolimeraseRESUMO
The construct containing the structural gene of the type II DNA topoisomerase of Drosophila melanogaster fused to the sequence of the chloroplast transit peptide directed by the 35 S promoter was produced on the basis of the binary vector Bin 19. The construct was introduced into Nicotiana tabacum plants by Agrobacterium-mediated transformation. The integration of the construct into genomic DNA was demonstrated by PCR and Southern blot hybridization. Expression of the chimeric gene at the transcription level was studied by Northern blotting. The protein produce of the expression was identified in chloroplasts by Western blot hybridization with polyclonal anti-type II Drosophila topoisomerase antibodies.
